EP2758080B1 - Immunothérapie anticancéreuse - Google Patents

Immunothérapie anticancéreuse Download PDF

Info

Publication number
EP2758080B1
EP2758080B1 EP12834327.4A EP12834327A EP2758080B1 EP 2758080 B1 EP2758080 B1 EP 2758080B1 EP 12834327 A EP12834327 A EP 12834327A EP 2758080 B1 EP2758080 B1 EP 2758080B1
Authority
EP
European Patent Office
Prior art keywords
tumor
tegvax
cells
gvax
agonist
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Not-in-force
Application number
EP12834327.4A
Other languages
German (de)
English (en)
Other versions
EP2758080A4 (fr
EP2758080A1 (fr
Inventor
Young Jun Kim
Drew M. Pardoll
Juan FU
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Johns Hopkins University
Original Assignee
Johns Hopkins University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Johns Hopkins University filed Critical Johns Hopkins University
Publication of EP2758080A1 publication Critical patent/EP2758080A1/fr
Publication of EP2758080A4 publication Critical patent/EP2758080A4/fr
Application granted granted Critical
Publication of EP2758080B1 publication Critical patent/EP2758080B1/fr
Not-in-force legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/13—Tumour cells, irrespective of tissue of origin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2818—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD28 or CD152
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/515—Animal cells
    • A61K2039/5152—Tumor cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55516—Proteins; Peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/80—Vaccine for a specifically defined cancer
    • A61K2039/876—Skin, melanoma

Definitions

  • This invention is related to the area of cancer therapy. In particular, it relates to cancer immunotherapy.
  • GVAX lethally irradiated tumor cell vaccines engineered to secrete GM-CSF
  • APC antigen presenting cells
  • TLR agonists One simple strategy that phenocopies the robust immunological responses seen in vaccines against infectious agents is to combine multiple TLR agonists with a cancer cell-based vaccine. Clinically, multiple adjuvants have been developed for cancer patients to augment the potency of cancer vaccines, and many of these adjuvants are typically TLR agonists.
  • TLR4 receptors expressed on tumor cells have shown to promote carcinogenesis.
  • TLR signaling in the hematopoietic compartment has been shown to elicit anti-tumor responses, which have translated into multiple clinical trials.
  • Li et al. (Clin. Cancer Res., 15, 1623 - 1634, 2009 ) discloses that the combination of PD-1 blockade with GM-CSF-secreting tumor cell immunotherapy leads to significantly improved antitumor responses by augmenting the tumor-reactive T-cell responses induced by the cellular immunotherapy.
  • composition which may be used for treating cancer patients.
  • the composition is as defined in appended claim 1.
  • composition of the invention is for administration to a cancer patient.
  • cancers include colorectal cancer, an aero-digestive squamous cancer, a lung cancer, a brain cancer, a liver cancer, a stomach cancer, a sarcoma, a leukemia, a lymphoma, a multiple myeloma, head-and-neck cancer, an ovarian cancer, cervical cancer, a uterine cancer, a breast cancer, a melanoma, a prostate cancer, a pancreatic carcinoma, and a renal carcinoma.
  • This list is meant to be illustrative rather than limiting.
  • Whole cancer cells may be allogeneic, syngeneic, or autologous to the treatment recipient. Typically they may be treated to make them proliferation incompetent by a technique which preserves preserve their immunogenicity and their metabolic activity. One typically used technique is irradiation. Such cells. Typically the same general type of tumor cell is used that the patient bears. For example, a patient suffering from melanoma will typically be administered proliferation incompetent melanoma cells.
  • the cells may express and secrete GM-CSF naturally or by transfection with a nucleic acid which directs such expression and secretion.
  • the tumor cell may express a transgene encoding GM-CSF as described in U.S. Pat. Nos.
  • Granulocyte-macrophage colony stimulating factor (GM-CSF) polypeptide is a cytokine or fragment having immunomodulatory activity and having at least about 85% amino acid sequence identity to GenBank Accession No. AAA52122.1.
  • Antibodies which are suitable for use in the treatment regimen and compositions and kits include any which specifically bind to Programmed Death 1 (PD-1).
  • PD-1 Programmed Death 1
  • Exemplary types of antibodies which may be employed include without limitation human, humanized, chimeric, monolclonal, polyclonal, single chain, antibody binding fragments, and diabodies.
  • antibodies are substantially encoded by an immunoglobulin gene or immunoglobulin genes, or fragments thereof.
  • Antibodies are capable of specifically binding an antigen or epitope. See, e.g. Fundamental Immunology, 3rd Edition, W.E. Paul, ed., Raven Press, N.Y. (1993 ); Wilson (1994; J. Immunol. Methods 175:267-273 ; Yarmush (1992) J. Biochem.
  • An antibody typically specifically binds to an antigen or epitope. Specific binding occurs to the corresponding antigen or epitope even in the presence of a heterogeneous population of proteins and other biologies. Specific binding of an antibody indicates that it binds to its target antigen or epitope with an affinity that is substantially greater than binding to irrelevant antigens
  • the relative difference in affinity is often at least 25% greater, more often at least 50% greater, most often at least 100%.
  • the relative difference can be at least 2x, at least 5x, at least 10x, at least 25x, at least 50x, at least 100x, at least 1000x, for example.
  • TLR Toll like receptors
  • DC dendritic cell
  • TLRs are conserved membrane spanning molecules containing an ectodomain of leucine-rich repeats, a transmembrane domain and an intracellular TIR. (Toll/IL-1R) domain.
  • TLRs recognize distinct structures in microbes, often referred to as "PAMPs" (pathogen associated molecular patterns).
  • PAMPs pathogen associated molecular patterns
  • Exemplary agonists which may be used for these receptors include, without limitation lipoproteins, lipopolypeptides, peptidoglycans, zymosan, lipopolysaccharide, neisserial porins, flagellin, profillin, galactoceramide, muramyl dipeptide, glucopyranosyl lipid A (GLA), and resiquimod (R848).
  • Peptidoglycans, lipoproteins, and lipoteichoic acids are cell wall components of Gram-positive. Lipopolysaccharides are expressed by most bacteria.
  • Flagellin is the structural component of bacterial flagella that is secreted by pathogenic and commensal bacterial.
  • a Galactosylceramide ( ⁇ -GalCer) is an activator of natural killer T (NKT) cells.
  • Muramyl dipeptide is a bioactive peptidoglycan motif common to all bacteria.
  • Such agonists mediate innate immune activation via Toll-like Receptors.
  • Specific binding of an agonist for its cognate receptor is often expressed in terms of an affinity.
  • K d k off /k on
  • TLRs that are expressed on the surface of cells include TLR-1,-2,-4,-5, and -6, while TLR-3, -7/8, and -9 are expressed with the ER compartment.
  • Human dendritic cell subsets can be identified on the basis of distinct TLR expression patterns.
  • the myeloid or "conventional" subset of DC expresses TLRs 1-8 when stimulated, and a cascade of activation markers (e.g. CD80, CD86, MHC class I and II, CCR7), pro-inflammatory cytokines, and chemokines are produced.
  • a cascade of activation markers e.g. CD80, CD86, MHC class I and II, CCR7
  • DCs acquire an enhanced capacity to take up antigens and present them in an appropriate form to T cells.
  • plasmacytoid subset of DC expresses only TLR7 and TLR9 upon activation, with a resulting activation of NK cells as well as T-cells.
  • activating DC with TLR agonists may be beneficial for priming anti-tumor immunity in an immunotherapy approach to the treatment of cancer. It has also been suggested that successful treatment of breast cancer using radiation and chemotherapy requires TLR4 activation.
  • TLR agonists known in the art and useful in the present invention include, but are not limited to, the following:
  • Formulation of the whole cancer cells with the TLR agonist appears to be a contributing factor to enhanced efficacy.
  • Formulations can be incubated together for periods of times such as 1 ⁇ 4, 1 ⁇ 2, 1, 2, 3, 5, 10, 24 hours, at temperatures such as 4 degrees C.
  • binding in the presence of a lipophilic agent or an emulsifying agent can be employed.
  • Such agents are well known in the art.
  • Various dosing schedules may be envisioned, with simultaneous or staggered timing, with single or multiple agents, single cycle or multiple cycles.
  • Methods of administering treatment agents to cancer patients vary. Exemplary methods include without limitation subcutaneous, intravenous, intramuscular, intraarterial, intradermal, intrathecal, intratumoral, intraperitoneal, sublingual, and epidural administrations. Administration may be to a human, mammal, mammalian subject, animal, veterinary subject, placebo subject, research subject, or experimental subject. Typically an agent such as an exogenous ligand, reagent, placebo, small molecule, pharmaceutical agent, therapeutic agent, diagnostic agent, or composition is contacted with the subject in an appropriate anatomical location. Administration may be for the purposes of therapy, pharmacokinetic study, diagnostic assay, research, placebo, or experimental method.
  • Agents according to the invention may be, but not need not be, administered as a single composition.
  • administration as a single composition is contemplated by the present invention, agents may be delivered to a single subject as separate administrations, which may be at the same or different time, and which may be by the same route or different routes of administration. In some cases, the agents may in fact contact each other within the subject's body, forming a composition in vivo.
  • GLA-TLR4 agonist glucopyranosyl lipid A
  • TLR7/8 agonist resiquimod
  • R848 is a TLR7/8 agonist that was found to produce 50-100 fold cytokines response compared to imiquimod (TLR7 agonist), and this agent has also passed phase I trials for safety in patients.
  • TLR7 agonist TLR7 agonist
  • TLR Agonist Enhanced GVAX TLR Agonist Enhanced GVAX
  • mice and reagents 6-8 weeks old female C57BL/6, Balb/c, and C3H/HeOUJ mice (Jackson Lab) were housed according to the Johns Hopkins Hospital (JHH) Animal Care and Use Committee.
  • C57BL/6 MyD88 -/- TRIF -/- , and B6 (Cg) Rag2tml (Rag2 -/- ) mice were obtained from Drs. Franck Housseau and Fan Pan, respectively (JHH).
  • 1316 and B16 GVAX cells were cultured in RPMI1640 media containing 10% ⁇ FCS, penicillin (100U/ml) and streptomycin (100U/ml).
  • CD11c+ cells were isolated by anti-mouse CD11c microBeads (MACS, Miltenyi Biotec). CD4 depleting GK1.5 antibody and CD8 depleting 2.43 (Bio X Cell) at 200 ⁇ g/dose, were injected intraperitoneally every 2 days (total 5 times). Hybridoma expressing blocking anti-PD-1 antibody (clone G4) was obtained from Dr. Charles Drake (JHH).
  • Glucopyranosyl lipid A (GLA) at 1mg/ml and resiquimod (R848) at 0.2mg/ml resiquimod were prepared in 10% (w/v) squalene oil-in-water emulsion (Immune Design).
  • IDC-1005 is the mixture of 1mg/ml GLA and 0.2mg/ml R848 in emulsion vehicle. IDC-1005 was incubated with irradiated GVAX cells at 4 deg C for 0.5-2 hours prior to inoculation.
  • GVAX formulated with LDC-1005 is labeled as TEGVAX.
  • GLA and R848 without emulsion vehicles were absorbed into GVAX cells with Lipofectamine and washed 4 times to remove non-absorbed TLR agonists and transfectants.
  • Tumor treatment assay C57BL/6 mice were injected with 1-5x10 4 B16 in the footpads. Once palpable tumor developed (5-10 days), 100 ⁇ l of 10 6 B16 GVAX formulated with or without IDC-1005 were injected subcutaneously into the contralateral limb. The control groups were injected with vehicle. For all these experiments, 10 mice were used per group. C3H/HeOUJ mice and Balb/c mice were used with SCCFVII/SF cells and CT26 cells, respectively with comparable methods (28). Previously prepared irradiated 10 6 SCCFVH/SF-GVAX was used for SCCFVII model in a same manner as B16 (3).
  • CT26 cells transduced with GM-CSF were used as CT26 GVAX (11).
  • GM-CSF was titered to ensure that GM-CSF expression level ranged from 50-500ng/10 6 cells/24 hours. Tumor was measured daily.
  • GVAX were labeled with Qtracker 655 (Invitrogen) prior to inoculation.
  • 100 ⁇ g/mice/injection was injected intraperitoneally twice a week once tumor was palpable in conjunction with vaccine treatments.
  • DC activation assay Spleens and draining lymph nodes (DLN) from tumor-bearing or naive mice were harvested 3-7 days post-vaccine treatments. Crushed spleens were digested in media containing DNAse I (Roche) and Liberase Blendzyme 2 (20,000 Mandl U/ml) (Roche), DC-enriched populations were obtained by depleting CD3 + and CD19 + , and gated for CD11c + and B220 + . These were evaluated by a multicolored FACS analysis using CD80, CD86, CD40, and MHCII antibodies.
  • ELISPOT assay ELISPOT plates (MultiScreen HTS filter plate, Millipore) were coated with a mouse IFN- ⁇ Ab (MabTech) for 24 hours and T2kb cells were pulsed with 10 ⁇ g/ml of P15E (KSPWFTTL) peptide overnight. 10 6 splenic CD8 cells from spleen were plated in triplicates to be co-cultured with pulsed or unpulsed 10 5 T2kb cells or stimulated with 1 ⁇ M of PMA and 10ng/ml of Ionomycin as positive controls. On day 3, biotinylated anti-mouse IFN- ⁇ Ab (MabTech) and Strepavidin-HRP were added. AEC Substrate Reagent (BD) was used to develop spots and analyzed using ELISPOT Plate Reader (Immunospot).
  • BD ELISPOT Plate Reader
  • In vivo CTL assay Splenocytes were labeled with 0.5 ⁇ M and 5 ⁇ M CFSE (Molecular Probes). The 5 ⁇ M CFSE labeled cells were pulsed with 10 ⁇ g/ml P15E (KSPWFTTL) peptide. The 0.5 ⁇ M CFSE labeled cells pulsed with ⁇ -gal (TPHPARIGL) peptide. Mice were injected intravenously with a 1:1 mixture of these cells, and splenocytes were isolated after 24 hours and analyzed by flow cytometry. Antigen-specific killing was calculated using the following formula: (1-% of CFSE P15E /% of CFSE ⁇ -gal ) X 100.
  • Immunohistochemistry 10 ⁇ m thick frozen sections were fixed with acetone, and blocked with 1% BSA for 30 minutes at RT. For paraffin embedded tissue, the sections were fixed in 4% paraformaldehyde prior to 1% BSA blocking as note above. ⁇ CD4, ⁇ CD8, ⁇ CD86 FITC conjugates and ⁇ CD45 and ⁇ B7-H1 primary antibodies were incubated 1 hour and 4°C, Cy3 conjugate antibody was used as secondary antibody in some cases. DAPI was used as the nuclear counterstain. Positive cells in 10 randomly selected fields at x40 magnification were quantitated. Quantitation of positive staining was blindly performed (LH) after the IHC slides were stained and marked randomly (JF). The microscope was Nikon, Eclipse E800. The camera was Nikon, DS-QilMc. The software was NIS-Element AR 3.0.
  • Cytokines analysis Harvested DC was cultured with GolgistopTM (BD) protein transport monensin and LPS 0.1 ⁇ g/ml 5hrs. DC was stained for anti-mouse CD11c, CD86, MHCII, IL-12, IFN ⁇ and TNF ⁇ expression, and harvested lymphocytes were stained for anti-mouse IL-2, IFN ⁇ , IFN ⁇ , TNF ⁇ expression after membrane permeabilization with Cytokit. Data were acquired by FACS analysis after rinsing out the antibodies.
  • GolgistopTM GolgistopTM
  • TLR agonists enhanced GVAX increased both activated conventional and plasmacytoid dendritic cells in the draining lymph nodes in comparison to GVAX
  • TEGVAX was able to enhance the activation phenotype of dendritic cells from the DLN of the vaccine inoculation site ( Figure 1 ). Both plasmacytoid DCs (pDC) and the conventional DCs (cDC) were analyzed, and both population showed enhanced expression of CD80 and CD86 activation markers for the TEGVAX treated group ( Figure 1A-C ).
  • Gated DCs showed increased activation markers peaking on day 3 after adjuvant injection and this persisted until day 7 ( Figure 1D ).
  • cytokine profiles from CD11c+ cells from the draining lymph nodes also demonstrated increase amount of IL-12, IFN ⁇ , and TNF ⁇ that can skew the T-cell repertoire towards T H 1 response (Supplemental Figure 1 ).
  • TEGVAX treatment of established tumors significantly reduced tumor growth rate in vivo.
  • mice that received TEGVAX displayed significantly lower tumor growth rate with only a single treatment. Both GVAX alone and TLR agonists alone had some modest benefit, but the combination treatment produced the best in vivo anti-tumor response.
  • TEGVAX with GVAX formulation with GLA alone or with R848 alone, we found that the combined GLA/R848 formulation into GVAX had the best anti-tumor response (Supplemental Figure 2A ).
  • TEGVAX treated mice did not display the exponential growth seen in untreated mice ( Figure 2B ).
  • TEGVAX was injected every 7 days because of the decreased level of activated DC at this point ( Figure 1D ).
  • These TEGVAX treated mice with smoldering tumors were inoculated with 10 5 B16 cells at a different site from the primary tumor site, and, in these mice, no tumor grew at the second site, demonstrating in vivo immunity against subsequent B16 challenges (data not shown).
  • TEGVAX The anti-tumor effect of TEGVAX was dependent on the formulation of the TEGVAX.
  • TEGVAX When we treated the tumor bearing mice with GVAX and GLA/R848 without 1 hour of co-incubation prior to injection, no anti-tumor effect was noted ( Fig. 2D ).
  • the co-incubation time can allow the hydrophobic TLR agonists to be absorbed into the GVAX with the squalene oil emulsion vehicle, and that this formulation is necessary for the anti-tumor response of TEGVAX.
  • TEGVAX generated from transfection method is comparable to the TEGVAX generated from prolonged incubation method with lipophilic vehicle in terms of their anti-tumor efficacy (Supplemental Figure 4 ).
  • TEGVAX increased lymphocytic infiltration in the tumor microenvironment, and this was associated with induction of tumor necrosis, locoregional T H 1 response, and increased B7-H1 expression on the tumor.
  • TEGVAX also significantly increased the expression of B7-H1 in comparison to GVAX and vehicle treated mice ( Figure 4C ).
  • TEGVAX's anti-tumor response was dependent on both CD4 and CD8 cells, as well as MyD88/TRIF signaling.
  • TEGVAX Since multiple TLR agonists were used as the formulation component for TEGVAX, we sought to determine whether TLR signaling was critical for the vaccine effect. TEGVAX treatments of B16 tumor-bearing mice were performed with MyD88/TRIF double knockout mice. Once again, the anti-tumor effect of TEGVAX was completely abrogated in these mice, confirming that TEGVAX's antitumor effect is dependent on TLR signaling ( Figure 5D ).
  • TEGVAX increases the number of tumor specific p15E specific cytotoxic T-cells
  • ELISPOT assays were also performed from CD8 T-cells from each of the treated and control groups with T2kb cells pulsed with p15E peptide as the APC. These two independent assays demonstrated increased numbers and activity of tumor specific (anti-p15E) CTLs in the mice treated with TEGVAX ( Figure 6B ).
  • TEGVAX combined with anti-PD-1 antibody can induce regression of established B16 tumor
  • TEGVAX Because of TEGVAX's ability to increase the number of p15E-specific cytotoxic T-cells as well as IFN ⁇ producing CD8 cells, we combined TEGVAX with the blockade of immune checkpoint pathway B7-H1/PD-1. PD-1 is expressed on activated T-cells and the co-localization of its ligand B7-H1, which is upregulated with IFN ⁇ secreting T-cells, on the tumor cells is correlated with clinical efficacy of anti-PD-1 blockade. The implication of these findings that supported the adaptive immune resistance mechanism is that anti-PD-1 blockade is ideally suited to be combined with a vaccine that can generate a powerful T H 1 response.
  • TEGVAX increased the both IFN ⁇ secreting T-cells as wells as upregulated the expression of B7-H1 on the tumor cells ( Figure 4 ).
  • anti-PD-1 antibody treatment was combined with TEGVAX, established B16 tumors were noted to regress in 50% of the mice treated ( Figure 7A ).
  • vitiligo was noted as shown in Figure 7B .
  • GLA and R848 were initially selected because these adjuvants significantly induced enhanced anti-tumor cytokine profiles compared to other TLR agonists, and both have been tested in patients to be safe.
  • GLA a TLR4 agonist
  • MPL monophosphoryl lipid A
  • R848, a TLR7/8 agonist was also found have significant improvements in increasing type I IFN profile over imiquimod. Our in vivo results on established tumors, therefore, demonstrated the high translational potential of TEGVAX as a cancer vaccine.
  • TIL Tumor infiltrating lymphocyte
  • ipilimumab is associated with grade 3-4 toxicity and isolated reports of therapy related mortality, so the addition of anti-CTLA-4 antibody may not be the first option to improve upon our in vivo results.
  • anti-CTLA-4 antibody may not be the first option to improve upon our in vivo results.
  • anti-PD-1 because of significant redundancy in the many families of immune checkpoint molecules as well as the availability of clinical grade checkpoint blocking antibodies, there are potentials to add other immune checkpoint blocking antibodies to anti-PD-1 treatment. Because of the likelihood that increased IFN ⁇ can potentially upregulate B7-H1 in the tumor microenvironment, an alternative approach would be to increase the level of tumor specific CTLs using a safe vaccine that can be combined with anti-PD-1.
  • TEGVAX TLR4 and TLR7/8 activity
  • TLR4 and TLR7/8 activity TLR4 and TLR7/8 activity
  • other TLR agonists Flagellin, CpG, etc
  • PAMP molecules PAMP molecules
  • TEGVAX treatment assay was performed with MyD88-TRIF double knockout mice, and the anti-tumor effect was completely abrogated ( Figure 4C ).
  • MyD88 is the critical mediator for TLR7/8 signaling
  • both TRIF and MyD88 is the critical downstream mediator of TLR4 signaling.
  • TEGVAX with Q-dot, and quantitated the endogenous APC's that was labeled via cell-to-cell transfer with Q-dot in the draining lymph node (Supplemental Figure 5 ).
  • TEGVAX offers a significant improvement over GVAX as a therapeutic vaccine that can significantly augment the priming of tumor specific T-cells, even for established tumors, and it is an excellent candidate to be added in a combinatorial therapy with anti-PD-1 blocking antibody or other forms of immune checkpoint blockade in cancer patients.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Immunology (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Cell Biology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oncology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Developmental Biology & Embryology (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biochemistry (AREA)
  • Zoology (AREA)
  • Biophysics (AREA)
  • Biotechnology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Hematology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicinal Preparation (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Claims (12)

  1. Composition comprenant :
    des cellules cancéreuses entières, incompétentes pour la prolifération, exprimant GM-CSF (facteur de stimulation des granulocytes et des monocytes) ;
    un anticorps anti-PD-1 qui se lie spécifiquement à Programmed Death 1 (PD-1) humaine ; et
    un agoniste de TLR4 (récepteur de type toll 4) et un agoniste de TLR7/8 (récepteur de type toll 7/8),
    où les cellules cancéreuses entières sont formulées avec les agonistes de TLR4 et TLR7/8.
  2. Composition selon la revendication 1 où l'agoniste de TLR4 est GLA.
  3. Composition selon la revendication 1 où l'agoniste de TLR7/8 est R848.
  4. Composition selon la revendication 1 où les cellules cancéreuses sont des cellules de mélanome.
  5. Composition selon la revendication 1 où l'agoniste de TLR et les cellules tumorales entières sont formulés avec un véhicule en émulsion.
  6. Composition selon la revendication 1 où l'agoniste de TLR et les cellules tumorales entières sont formulés avec de la Lipofectamine™.
  7. Composition selon la revendication 1 où l'agoniste de TLR et les cellules tumorales entières sont formulés avec un lipide cationique.
  8. Composition selon l'une quelconque des revendications 1 à 7 destinée à être utilisée dans le traitement d'un patient cancéreux.
  9. Composition destinée à être utilisée selon la revendication 8, où les cellules cancéreuses entières sont autologues pour le patient.
  10. Kit comprenant les agents :
    des cellules cancéreuses entières, incompétentes pour la prolifération, exprimant GM-CSF (facteur de stimulation des granulocytes et des monocytes) ;
    un anticorps anti-PD-1 qui se lie spécifiquement à Programmed Death 1 (PD-1) humaine ; et
    un agoniste de TLR4 (récepteur de type toll 4) et un agoniste de TLR7/8 (récepteur de type toll 7/8).
  11. Kit selon la revendication 10 comprenant en outre des instructions pour administrer et/ou formuler les agents.
  12. Kit selon la revendication 10 comprenant en outre :
    (1) un véhicule en émulsion ;
    (2) de la Lipofectamine™ ; ou
    (3) un lipide cationique.
EP12834327.4A 2011-09-19 2012-09-19 Immunothérapie anticancéreuse Not-in-force EP2758080B1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201161536115P 2011-09-19 2011-09-19
PCT/US2012/055992 WO2013043647A1 (fr) 2011-09-19 2012-09-19 Immunothérapie anticancéreuse

Publications (3)

Publication Number Publication Date
EP2758080A1 EP2758080A1 (fr) 2014-07-30
EP2758080A4 EP2758080A4 (fr) 2015-03-11
EP2758080B1 true EP2758080B1 (fr) 2018-03-07

Family

ID=47914812

Family Applications (1)

Application Number Title Priority Date Filing Date
EP12834327.4A Not-in-force EP2758080B1 (fr) 2011-09-19 2012-09-19 Immunothérapie anticancéreuse

Country Status (7)

Country Link
US (1) US9308253B2 (fr)
EP (1) EP2758080B1 (fr)
JP (2) JP2014527983A (fr)
CN (1) CN103957939A (fr)
CA (1) CA2849259A1 (fr)
HK (1) HK1197182A1 (fr)
WO (1) WO2013043647A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10837018B2 (en) 2013-07-25 2020-11-17 Exicure, Inc. Spherical nucleic acid-based constructs as immunostimulatory agents for prophylactic and therapeutic use

Families Citing this family (38)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011133636A1 (fr) 2010-04-20 2011-10-27 Cedars-Sinai Medical Center Polythérapie mettant en œuvre la déplétion en lymphocytes cd4 et inhibiteurs de mtor
WO2011136828A1 (fr) * 2010-04-27 2011-11-03 The Johns Hopkins University Compositions immunogènes et méthodes de traitement d'une néoplasie
CN112587671A (zh) 2012-07-18 2021-04-02 博笛生物科技有限公司 癌症的靶向免疫治疗
SG10201710472PA (en) * 2013-02-22 2018-02-27 Curevac Ag Combination of vaccination and inhibition of the pd-1 pathway
CN104415335A (zh) * 2013-09-02 2015-03-18 北京中康万达医药科技有限公司 体内个体化***免疫治疗方法和装置
EP3757130A1 (fr) 2013-09-26 2020-12-30 Costim Pharmaceuticals Inc. Méthodes de traitement de cancers hématologiques
US10241115B2 (en) 2013-12-10 2019-03-26 Merck Sharp & Dohme Corp. Immunohistochemical proximity assay for PD-1 positive cells and PD-ligand positive cells in tumor tissue
DK3081576T3 (da) 2013-12-12 2019-10-21 Shanghai hengrui pharmaceutical co ltd Pd-1-antistof, antigenbindende fragment deraf og medicinsk anvendelse deraf
CN114716555A (zh) 2014-01-10 2022-07-08 博笛生物科技有限公司 靶向表达egfr的肿瘤的化合物和组合物
JOP20200094A1 (ar) 2014-01-24 2017-06-16 Dana Farber Cancer Inst Inc جزيئات جسم مضاد لـ pd-1 واستخداماتها
JOP20200096A1 (ar) 2014-01-31 2017-06-16 Children’S Medical Center Corp جزيئات جسم مضاد لـ tim-3 واستخداماتها
JP2017507931A (ja) * 2014-02-05 2017-03-23 シーダーズ−サイナイ メディカル センター がん及び感染症の治療方法並びに治療用組成物
KR102290205B1 (ko) 2014-06-04 2021-08-20 엑시큐어, 인크. 예방 또는 치료 용도를 위한 리포솜성 구형 핵산에 의한 면역 조절인자의 다가 전달
CN105233291A (zh) * 2014-07-09 2016-01-13 博笛生物科技有限公司 用于治疗癌症的联合治疗组合物和联合治疗方法
AU2015286043B2 (en) * 2014-07-09 2020-08-20 Birdie Biopharmaceuticals Inc. Anti-PD-L1 combinations for treating tumors
CN112587672A (zh) * 2014-09-01 2021-04-02 博笛生物科技有限公司 用于***的抗-pd-l1结合物
EP3925622A1 (fr) 2014-09-13 2021-12-22 Novartis AG Polythérapies
TWI716362B (zh) 2014-10-14 2021-01-21 瑞士商諾華公司 針對ｐｄ－ｌ１之抗體分子及其用途
WO2017004421A1 (fr) * 2015-06-30 2017-01-05 The Trustees Of The University Of Pennsylvania Compositions topiques et injectables de résiquimod pour le traitement d'affections cutanées néoplasiques
US20180222982A1 (en) * 2015-07-29 2018-08-09 Novartis Ag Combination therapies comprising antibody molecules to pd-1
WO2017024296A1 (fr) * 2015-08-06 2017-02-09 Memorial Sloan Kettering Cancer Center Procédés et compositions destinés à la thérapie de tumeurs
US20180303845A1 (en) * 2015-11-02 2018-10-25 Ventirx Pharmaceuticals, Inc. Use of tlr8 agonists to treat cancer
CN115350279A (zh) 2016-01-07 2022-11-18 博笛生物科技有限公司 用于***的抗-her2组合
CN115252792A (zh) 2016-01-07 2022-11-01 博笛生物科技有限公司 用于***的抗-egfr组合
CN115554406A (zh) 2016-01-07 2023-01-03 博笛生物科技有限公司 用于***的抗-cd20组合
KR20230149857A (ko) 2016-07-07 2023-10-27 더 보드 어브 트러스티스 어브 더 리랜드 스탠포드 주니어 유니버시티 항체-애쥬번트 접합체
JP7321489B2 (ja) * 2016-07-27 2023-08-07 株式会社先端免疫療法研究所 がん免疫アジュバント
WO2018039629A2 (fr) 2016-08-25 2018-03-01 Northwestern University Acides nucléiques sphériques micellaires obtenus à partir de matrices thermosensibles sans trace
CN110114057A (zh) * 2016-09-15 2019-08-09 艾德拉药物股份有限公司 Tlr9激动剂对癌症治疗的免疫调节作用
CN108794467A (zh) 2017-04-27 2018-11-13 博笛生物科技有限公司 2-氨基-喹啉衍生物
MX2019015744A (es) 2017-06-23 2020-02-20 Birdie Biopharmaceuticals Inc Composiciones farmaceuticas.
AU2018316694A1 (en) * 2017-08-17 2020-02-13 Nektar Therapeutics Immunotherapeutic tumor treatment method
US20200113986A1 (en) * 2018-10-10 2020-04-16 The Research Foundation Of The State University Of New York Genome edited cancer cell vaccines
EP3924521A4 (fr) 2019-02-15 2023-03-29 IncellDx, Inc. Dosage d'échantillons associés à la vessie, identification et traitement d'une néoplasie associée à la vessie, et kits destinés à être utilisés dans ces derniers
EP3937984A1 (fr) 2019-03-15 2022-01-19 Bolt Biotherapeutics, Inc. Immunoconjugués ciblant le her2
US20230022045A1 (en) * 2019-12-03 2023-01-26 Board Of Regents, The University Of Texas System Combination therapies for the treatment of cancer
CN110812476B (zh) * 2019-12-18 2022-07-26 福建医科大学附属协和医院 弥漫大b细胞淋巴瘤的免疫佐剂及其应用
WO2021199056A1 (fr) * 2020-04-03 2021-10-07 Kanishk Sinha Vaccin pour nouveau coronavirus

Family Cites Families (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5904920A (en) 1991-10-04 1999-05-18 Whitehead Institute For Biomedical Research Regulation of systemic immune responses utilizing cytokines and antigens
US5637483A (en) 1991-10-04 1997-06-10 Whitehead Institute For Biomedical Research Irradiated tumor cell vaccine engineered to express GM-CSF
US6033674A (en) 1995-12-28 2000-03-07 Johns Hopkins University School Of Medicine Method of treating cancer with a tumor cell line having modified cytokine expression
US6277368B1 (en) 1996-07-25 2001-08-21 The Regents Of The University Of California Cancer immunotherapy using autologous tumor cells combined with cells expressing a membrane cytokine
US20040197312A1 (en) * 2003-04-02 2004-10-07 Marina Moskalenko Cytokine-expressing cellular vaccine combinations
GB0321615D0 (en) * 2003-09-15 2003-10-15 Glaxo Group Ltd Improvements in vaccination
WO2007120368A2 (fr) 2006-01-09 2007-10-25 The Regents Of The University Of California Combinaisons immunostimulatrices de tnfrsf, tlr, nlr, rhr, du récepteur purinergique et des agonistes du récepteur de la cytokine utilisées pour les vaccins et en immunothérapie anti-tumorale
WO2009014708A2 (fr) * 2007-07-23 2009-01-29 Cell Genesys, Inc. Anticorps pd-1 en combinaison avec une cellule sécrétant de la cytokine et leurs procédés d'utilisation
CN102202689A (zh) * 2008-08-28 2011-09-28 葛兰素史密丝克莱恩生物有限公司 疫苗
US8840881B2 (en) * 2008-08-28 2014-09-23 Aduro Gvax Inc. Methods and compositions for treating prostate cancer or inducing a humoral immune response against prostate cancer
KR101050829B1 (ko) * 2008-10-02 2011-07-20 서울대학교산학협력단 항 ｐｄ-1 항체 또는 항 ｐｄ-ｌ1 항체를 포함하는 항암제
WO2010130897A1 (fr) * 2009-05-14 2010-11-18 Sanofi Pasteur Procédé de détoxification du lipopolysaccharide (lps) ou du lipide a des bactéries à gram-négatif
WO2011136828A1 (fr) * 2010-04-27 2011-11-03 The Johns Hopkins University Compositions immunogènes et méthodes de traitement d'une néoplasie

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
"Bioconjugate Techniques, 2nd Edition", 1 January 2008, ELSEVIER, article GREG T. HERMANSON: "Heterobifunctional PEG Reagents", pages: 718 - 719, XP055340009 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10837018B2 (en) 2013-07-25 2020-11-17 Exicure, Inc. Spherical nucleic acid-based constructs as immunostimulatory agents for prophylactic and therapeutic use

Also Published As

Publication number Publication date
CA2849259A1 (fr) 2013-03-28
US9308253B2 (en) 2016-04-12
HK1197182A1 (en) 2015-01-09
US20140341978A1 (en) 2014-11-20
EP2758080A4 (fr) 2015-03-11
EP2758080A1 (fr) 2014-07-30
JP2014527983A (ja) 2014-10-23
JP2018021047A (ja) 2018-02-08
WO2013043647A1 (fr) 2013-03-28
CN103957939A (zh) 2014-07-30

Similar Documents

Publication Publication Date Title
EP2758080B1 (fr) Immunothérapie anticancéreuse
Fu et al. Preclinical evidence that PD1 blockade cooperates with cancer vaccine TEGVAX to elicit regression of established tumors
Kumai et al. Optimization of peptide vaccines to induce robust antitumor CD4 T-cell responses
Cerundolo et al. Harnessing invariant NKT cells in vaccination strategies
JP7111384B2 (ja) 悪性腫瘍の治療に対する方法
Kaczanowska et al. TLR agonists: our best frenemy in cancer immunotherapy
Zaini et al. OX40 ligand expressed by DCs costimulates NKT and CD4+ Th cell antitumor immunity in mice
CN105209047B (zh) 用于癌症治疗的gla单一疗法
Grauer et al. Immunotherapy of diffuse gliomas: biological background, current status and future developments
Brandonisio et al. Dendritic cells in Leishmania infection
Sharma et al. Costimulation as a platform for the development of vaccines: a peptide-based vaccine containing a novel form of 4-1BB ligand eradicates established tumors
Fujii et al. Adjuvant activity mediated by iNKT cells
McWilliams et al. Multiple innate signaling pathways cooperate with CD40 to induce potent, CD70-dependent cellular immunity
CN102170908A (zh) 向个体的肿瘤引流***递送cd40激动剂
Yu et al. Chicken HSP70 DNA vaccine inhibits tumor growth in a canine cancer model
JP2022512161A (ja) 免疫療法のための組成物及び方法
EP2911683B1 (fr) Composition enrichie en autophagosome allogénique pour le traitement de maladies
Sharma et al. Systemic Targeting of CpG-ODN to the Tumor Microenvironment with Anti–neu-CpG Hybrid Molecule and T Regulatory Cell Depletion Induces Memory Responses in BALB-neuT Tolerant Mice
Yan et al. Regulatory T-cell depletion synergizes with gp96-mediated cellular responses and antitumor activity
Dong et al. Co-operation of α-galactosylceramide-loaded tumour cells and TLR9 agonists induce potent anti-tumour responses in a murine colon cancer model
Zhang et al. Bidirectional crosstalk between therapeutic cancer vaccines and the tumor microenvironment: Beyond tumor antigens
van den Engel et al. Combination immunotherapy and active-specific tumor cell vaccination augments anti-cancer immunity in a mouse model of gastric cancer
Tu et al. Incorporation of a TGF-β2-inhibiting oligodeoxynucleotide molecular adjuvant into a tumor cell lysate vaccine to enhance antiglioma immunity in mice
Wojas-Turek et al. Antitumor effect of murine dendritic and tumor cells transduced with IL-2 gene
Petley Assessment of combined vaccination and immune modulation as an anti-tumour therapy

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20140411

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

DAX Request for extension of the european patent (deleted)
REG Reference to a national code

Ref country code: HK

Ref legal event code: DE

Ref document number: 1197182

Country of ref document: HK

A4 Supplementary search report drawn up and despatched

Effective date: 20150211

RIC1 Information provided on ipc code assigned before grant

Ipc: A61P 35/00 20060101ALI20150205BHEP

Ipc: A61K 38/19 20060101ALI20150205BHEP

Ipc: A61K 39/395 20060101AFI20150205BHEP

Ipc: A61K 38/18 20060101ALI20150205BHEP

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: EXAMINATION IS IN PROGRESS

17Q First examination report despatched

Effective date: 20170203

GRAP Despatch of communication of intention to grant a patent

Free format text: ORIGINAL CODE: EPIDOSNIGR1

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: GRANT OF PATENT IS INTENDED

INTG Intention to grant announced

Effective date: 20170728

GRAJ Information related to disapproval of communication of intention to grant by the applicant or resumption of examination proceedings by the epo deleted

Free format text: ORIGINAL CODE: EPIDOSDIGR1

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: EXAMINATION IS IN PROGRESS

INTC Intention to grant announced (deleted)
GRAR Information related to intention to grant a patent recorded

Free format text: ORIGINAL CODE: EPIDOSNIGR71

GRAS Grant fee paid

Free format text: ORIGINAL CODE: EPIDOSNIGR3

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: GRANT OF PATENT IS INTENDED

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE PATENT HAS BEEN GRANTED

AK Designated contracting states

Kind code of ref document: B1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

INTG Intention to grant announced

Effective date: 20180129

REG Reference to a national code

Ref country code: GB

Ref legal event code: FG4D

REG Reference to a national code

Ref country code: CH

Ref legal event code: EP

Ref country code: AT

Ref legal event code: REF

Ref document number: 975842

Country of ref document: AT

Kind code of ref document: T

Effective date: 20180315

REG Reference to a national code

Ref country code: IE

Ref legal event code: FG4D

REG Reference to a national code

Ref country code: DE

Ref legal event code: R096

Ref document number: 602012043840

Country of ref document: DE

REG Reference to a national code

Ref country code: NL

Ref legal event code: MP

Effective date: 20180307

REG Reference to a national code

Ref country code: LT

Ref legal event code: MG4D

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: HR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

Ref country code: FI

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

Ref country code: NO

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180607

Ref country code: LT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

Ref country code: CY

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

Ref country code: ES

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

REG Reference to a national code

Ref country code: HK

Ref legal event code: GR

Ref document number: 1197182

Country of ref document: HK

REG Reference to a national code

Ref country code: AT

Ref legal event code: MK05

Ref document number: 975842

Country of ref document: AT

Kind code of ref document: T

Effective date: 20180307

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: LV

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

Ref country code: RS

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

Ref country code: SE

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

Ref country code: BG

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180607

Ref country code: GR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180608

REG Reference to a national code

Ref country code: FR

Ref legal event code: PLFP

Year of fee payment: 7

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: RO

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

Ref country code: AL

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

Ref country code: IT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

Ref country code: EE

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

Ref country code: PL

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

Ref country code: NL

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: DE

Payment date: 20180927

Year of fee payment: 7

Ref country code: FR

Payment date: 20180925

Year of fee payment: 7

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: SM

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

Ref country code: SK

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

Ref country code: CZ

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

Ref country code: AT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: GB

Payment date: 20180927

Year of fee payment: 7

REG Reference to a national code

Ref country code: DE

Ref legal event code: R097

Ref document number: 602012043840

Country of ref document: DE

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: PT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180709

PLBE No opposition filed within time limit

Free format text: ORIGINAL CODE: 0009261

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: DK

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

26N No opposition filed

Effective date: 20181210

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: SI

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: MC

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

REG Reference to a national code

Ref country code: CH

Ref legal event code: PL

REG Reference to a national code

Ref country code: BE

Ref legal event code: MM

Effective date: 20180930

REG Reference to a national code

Ref country code: IE

Ref legal event code: MM4A

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: LU

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20180919

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: IE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20180919

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: BE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20180930

Ref country code: CH

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20180930

Ref country code: LI

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20180930

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: MT

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20180919

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: TR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180307

REG Reference to a national code

Ref country code: DE

Ref legal event code: R119

Ref document number: 602012043840

Country of ref document: DE

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: HU

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT; INVALID AB INITIO

Effective date: 20120919

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: MK

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20180307

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: DE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20200401

Ref country code: IS

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 20180707

GBPC Gb: european patent ceased through non-payment of renewal fee

Effective date: 20190919

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: GB

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20190919

Ref country code: FR

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20190930